The morphological and growth characteristics, activities of seven selected enzymes (ADH, LDH, MDH, ME, ICDH, GPGDH, and G6PDH), and enzyme patterns of ADH of four strains of freshwater fish trypanosomes were assessed and also compared to those of the species Trypanosoma boissoni RANQUE, 1967 from a marine host. It was shown that the strain from marine fish displays clear differences and is well distinguishable from the strains isolated from freshwater fish. The culture forms of the latter strains in turn exhibit a considerable mutual similarity although they were isolated from different freshwater hosts. This similarity indicates that close relationships exist between freshwater fish trypanosomes.